Can you use a goat anti-rabbit secondary and a rabbit anti-goat
secondary in the same reaction? Every fibre of my being says no -
they'd either precipitate out or at the very least give nonspecific
signals. On the other hand, I'm reading a paper that claims to have
done just that.
To reiterate, they are trying to detect two proteins, A and B. The
primary ABs are rabbit anti-A and goat anti-B. The secondary ABs are
Cy5-[goat anti-rabbit IgG] and FITC-[rabbit anti-goat IgG] conjugates.
Surely these two secondaries should mutually detect each other and
obliterate any signal from the underlying A and B? They're claiming a
perfect non-overlap of signals, with 50% of cells containing A and 50%
containing B.
Someone here is missing something, and I really hope it's not me.
Peter Ellis.